Effects of sulfate and chloride on three separate oxalate transporters reconstituted from rabbit renal cortex.
Understanding the mechanism of sulfate-dependent, oxalate-stimulated chloride reabsorption in the mammalian proximal tubule is complicated by the presence of multiple oxalate and sulfate transport pathways. Accordingly, we developed a method of reconstituting functional oxalate transport from the rabbit renal cortex so that the individual transporters might be examined. Solubilized microvillus membrane proteins were separated by hydroxyapatite chromatography and then reconstituted into proteoliposomes. Two peaks of oxalate/oxalate exchange activity were observed. Sulfate (10 mM) cis-inhibits oxalate transport in the early peak by 93% and in the later peak by 41%. In contrast, 20 mM chloride inhibits oxalate/oxalate exchange by only 32% in the early peak but inhibits oxalate exchange by 70% in the later peak. Oxalate-stimulated sulfate uptake was observed in the early fractions but not in the later fractions. These data are consistent with the recovery of the sulfate/oxalate exchanger in the early hydroxyapatite fractions and the chloride/oxalate exchanger in the later fractions. The basolateral membrane sulfate/oxalate exchanger was also reconstituted. The reconstituted basolateral and apical membrane sulfate/oxalate exchangers demonstrate nearly identical patterns of substrate specificities. However, 98% of apical sulfate/oxalate exchange activity is lost following exposure to octylglucoside at room temperature, whereas the basolateral sulfate/oxalate exchange activity was reduced 67% (P < 0.05). In conclusion, functional reconstitution of solubilized membrane proteins demonstrates that apical membrane chloride/oxalate exchange and sulfate/oxalate exchange are mediated by different transport proteins. Apical and basolateral sulfate/oxalate exchange may also represent transport on two separate exchangers.
Karniski
L P
LP
Department of Internal Medicine, Iowa City, Veterans Affairs, Iowa City, USA.
D000818:Animals; D027321:Anion Transport Proteins; D017920:Antiporters / Q000378:metabolism*; D002712:Chlorides / Q000494:pharmacology*; D007672:Kidney Cortex / Q000378:metabolism*; D007700:Kinetics; D008297:Male; D008871:Microvilli / Q000378:metabolism*; D010070:Oxalates / Q000378:metabolism*; D011817:Rabbits; D000076988:Sulfate Transporters; D013431:Sulfates / Q000494:pharmacology*
D027321:Anion Transport Proteins; D017920:Antiporters; D002712:Chlorides; D010070:Oxalates; C056459:Slc26a1 protein, rat; D000076988:Sulfate Transporters; D013431:Sulfates